Veterinary clinical pathology最新文献

Background: The Grand Cayman blue iguana (Cyclura lewisi ) is an endangered species of lizard endemic to Grand Cayman Island. The cytochemical staining characteristics, cell morphometrics, and ultrastructure have not been reported for blood from this species.

Objectives: The objectives for this blue iguana blood cell study were to perform routine and cytochemical staining, document morphometric measurements, and identify structures using bright-field (BF), differential interference contrast (DIC), scanning electron microscopy (SEM), and transmission electron microscopy (TEM).

Methods: Opportunistic blood samples were collected from one captive adult male blue iguana housed at the Milwaukee County Zoo, Milwaukee, WI, USA, during three routine health exams. CBCs, biochemistry panels, cytochemical staining (leukocyte acid phosphatase [AcP] with and without tartrate, alkaline phosphatase [ALP], alpha-naphthyl acetate esterase [ANAE], leukocyte peroxidase [LEP], periodic acid-Schiff [PAS], Sudan black B [SBB], and Toluidine blue [TB]) were performed.

Results: CBCs and biochemistry panel results were within range for a healthy captive adult male blue iguana. The cytochemical reactions were determined. The basophils, erythrocytes, heterophils, monocytes, and thrombocytes reacted positively to PAS. Only the heterophils had positive reactions to ALP, SBB, and LEP. Basophils were positive to TB. Monocytes and thrombocytes stained positive for AcP. Red blood cell cytoplasmic inclusions held stain with ANAE, AcP, PAS, and TB. The morphology and ultrastructure of blue iguana blood cells were documented using BF, DIC, SEM, and TEM.

Conclusion: All of the microscopic and staining methods allowed for improved differentiation of cell types and characteristics. The results may serve as a reference for any study of blood cells in other reptile species.

Background: microRNAs (miRNA) have been proposed as biomarkers for pancreatitis in dogs due to their high peak concentrations and experimental correlation with acinar cell injury. However, analytical validation and the effect of interfering substances are unknown.

Objective: The study aimed to (i) develop and analyze the analytical validity of an assay for miRNA-148a and miRNA-375 in serum and (ii) compare miRNA-148a and miRNA-375 between healthy and pancreatitis-affected dogs.

Methods: Circulating miRNAs were quantified from serum of healthy (n = 40) and pancreatitis-affected dogs (n = 40). Reference intervals were established, and serum miRNAs were compared between groups. Linearity was assessed via dilutional parallelism (observed/expected ratio, O/E). Precision was assessed via intra- and inter-assay coefficients of variation (% CV). Intralipid, bilirubin, and hemoglobin were tested as interferants.

Results: For miRNA-148a, the RI was < 33 000 gene copies and mean O/E was 100.4%. Mean inter-assay and intra-assay % CV were 1.5% and 1.3% respectively. For miRNA-375, the RI was < 4 gene copies and mean O/E was 100.1%. Mean inter-assay and intra-assay % CV were 3.2% and 0.5% respectively. Hemolysis led to false increases in both assays (p = 0.003 - < 0.0001). miRNA-148a was reduced in dogs with pancreatitis (mean 192 gene copies) compared with healthy controls (mean 2760 gene copies; p < 0.0001), but miRNA-375 was not different between groups (p = 0.94).

Conclusions: The miRNA assays were analytically valid; however, hemolysis, a common interferant in pancreatitis cases, may impact test results. miRNA-148a was suppressed in dogs with pancreatitis. It is unclear whether this represents a cause or a consequence of the disease.

Background: Hemorheology is used in human medicine but remains largely unexplored in veterinary medicine. The assessment of RBC rheology holds potential for diagnosing, monitoring, and predicting outcomes in veterinary species. Reference intervals (RIs) are needed before clinical assessment of the method.

Objectives: Determine RIs for hemorheological parameters in dogs, compare RIs generated at 620 versus 800 nm wavelengths, as well as RIs generated with and without following ASVCP guidelines.

Methods: Whole blood was sampled on EDTA tubes from healthy dogs and run on the MIZAR Analyzer for 13 parameters (DI, Int (d), PEAK, AI, Int (a), AMP, BpL, Bp, BpH, T½, OTF, OT, ED). After outlier removal considerations, the normality of distribution was assessed, and RIs were calculated using parametric or non-parametric methods (ASVCP guidelines). RIs performed at 620 and 800 nm, with and without following ASVCP guidelines, were compared.

Results: RIs were established for rheology parameters at 620 and 800 nm in a population of 42 healthy dogs. 5/13 parameters had nearly identical RI limits between 620 and 800 nm; others showed an overt shift at 800 nm toward higher values for both RI limits. Reference intervals showed similar results whether ASVCP guidelines were followed or not for 7/13 parameters, namely when data had a Gaussian distribution and contained a maximum of one outlier. For the remaining parameters, RIs calculated without following ASVCP guidelines differed significantly and were considered unsuitable for clinical use.

Conclusions: Specific RIs were established for 620 and 800 nm wavelengths. Compliance with ASVCP guidelines was deemed necessary.

Background: Point-of-care testing (POC) is widely utilized for rapid results for many different analytes. A new feline-specific N-terminal pro-brain natriuretic peptide (NT-proBNP) quantitative assay (Vcheck V200, Bionote Inc) is currently available but has not undergone independent validation.

Objective: To validate the Vcheck POC quantitative assay for feline NT-proBNP.

Methods: Validation was performed in accordance with the American Society for Veterinary Clinical Pathology guidelines utilizing serum samples from 62 cats. Precision was determined for low (50-100 pmol/L), mid (101-300 pmol/L), and high (> 301 pmol/L) pools short-term (20 repetitions) and long-term (5 repetitions each day for 5 days). Linearity, methods comparison, interference testing, and sample stability were evaluated.

Results: The within-day coefficients of variability (CV) were low pool = 12.6%, mid pool = 10.4%, and high pool = 8.7%. The within-week CV was low pool = 9.9%, mid pool = 14.9%, and high pool = 6.9%. The assay was linear over the analytical range of 53-1488 pmol/L (R² = 0.99, p < 0.0001). Paired samples between the feline Cardiopet NT-proBNP (IDEXX) and Vcheck assays demonstrated a mean difference of 11 pmol/L (2.3%), p = 0.38, between assays with minimal constant or proportional bias. Hemolysis and lipemia did not affect assay performance, while all icteric samples were invalid. Significantly lower values were identified in samples after 2 and 4 h when stored at 20°C and 4°C, respectively.

Conclusions: The Vcheck V200 has acceptable precision, accuracy, and compares favorably with commercially available assays and is a viable POC quantitative assay for feline NT-proBNP.

Background: The ProCyte One is a veterinary hematology analyzer. It uses laser flow cytometry with four optical detectors to categorize RBCs, WBCs, and platelets.

Objective: We aimed to compare results from the ProCyte One with the ProCyte Dx and ADVIA 120, as well as the results of manual PCVs and WBC differential counts, to determine their performance for automated CBCs in dogs and cats.

Materials and methods: ProCyte One inter-assay and intra-assay precision were assessed using control material and patient samples. For analyzer comparison, patient samples were run on all three analyzers within 2 h of each other. Passing-Bablok regression and visual inspection of Bland-Altman plots were used to determine analyzer congruence, and Spearman's rank correlation was performed.

Results: Precision was generally excellent, with CV < 4% for most measurands except for reticulocytes, lymphocytes, monocytes, and eosinophils. A total of 140 canine and 96 feline samples were included for the comparison study. Correlations were ≥ 0.93 for all comparisons except mean cell volume (MCV), mean cell hemoglobin concentration (MCHC), reticulocytes, lymphocytes, monocytes, and eosinophils. Statistically significant bias was suggested by Passing-Bablok regression for many measurands, but most were minor except for MCV compared to ADVIA 120, monocytes compared to all methods, and RBC/hematocrit in anemic dogs.

Conclusions: The ProCyte One demonstrates acceptable agreement with ProCyte Dx and ADVIA 120. However, because of the potential for significant differences between methods, operators should exercise caution when comparing patient results between different analyzers.